Extracts of gelling agents have been shown to stimulate rooting and this study was initiated to investigate the presence of root stimulating substances in gelling agents. After screening a number of gelling agents, four were selected, namely; Agar Bacteriological, Agar Commercial Gel, Difco Bacto Agar and Gelrite were selected and examined for the presence of root-stimulating substances using mungbean bioassay. Water extracts of Agar Bacteriological, Agar Commercial Gel and Difco Bactol Agar stimulated rooting of mungbean cuttings. Addition of Charcoal neither reduced nor increased rooting produced by the water extract of the first two agars but when added in conjunction with Difco Bacto Agar rooting was reduced. Autoclaving, however reduced rooting in extracts of the gelling agents. The possibility that root-stimulating substances may not be the same in all the gelling agents can not be excluded. Extraction of Gelrite with water was problematical and was therefore excluded. IBA solution and water extracts of the gelling agents separately promoted good rooting in mungbeans cuttings. Rooting in extracts of autoclaved frozen-thawed gelling agents was poor, however, IBA + gelling agents gave high rooting at the 100% concentration and this could possibly be due to an additive effect of the IBA. Addition of charcoal reduced rooting significantly in extracts of IBA + gelling agents. Using 80% acidic methanol, reasonable levels of rooting substances were obtained from the residue extract of this complex (IBA + gelling agent+charcoal) of all the gelling agents except Gelrite indicating that root-promoting substances were adsorbed by charcoal. The low rooting in the presence of the Gelrite extract was attributed to the matrix of the polymer of the Gelrite. Ethyl acetate fractionated extracts (EA-pH 8.0; EA-pH 3.0; and Aqueous) obtained from the four gelling agents stimulated rooting indicating the presence of numerous root promoting substances. Gelrite gave good rooting with both the 50 and 100% concentrations of all the fractions. Purified water and ethanol extracts of the gelling agents exhibited auxin-like activity when separated by paper chromatography and compared with IBA and IAA standards. Using HPLC, IAA was quantified in all the gelling agents with Difco Bacto Agar and Agar Commercial Gel having the highest IAA concentration and Gelrite the lowest IAA concentration. IAA concentration in Agar Bacteriological was a third of the level detected in Difco Bacto Agar. The information from this work may enable researchers to consider gelling agents as sources of auxin-like compounds and other plant hormones as well as support media for use in tissue culture procedures and also increase the enthuse for further research into the nutrient types and levels in gelling agents. / Thesis (Ph.D.)-University of Natal, Pietermaritzburg, 2003.

The purpose of the study was to determine how the waste from m-Endo agar LES (LesEndo) should be categorized and managed, due to its content of the carcinogenic substance basic red 9 in basic fuchsine. The information was obtained by studying legislation and a laboratory practical was performed to verify, if basic fuchsine could remain in LesEndo agar after being heated. This was important to know for the further interpretation how to categorize and manage the waste according to the legislation. LesEndo agar was autoclaved at 121 °C for 15 and 30 minutes. E. coli and coliform bacteria was put to grow on the LesEndo agar. The results from the study showed that all replicas had coliform colonies, which appeared with a metallic fuchsine-sheen. This verified that basic fuchsine was unchanged and that there was no significant difference in growth between replicas with heated agar and the control. The total concentration of basic fuchsine in LesEndo agar was 0.08%. To be categorized as hazardous waste the threshold value for basic red 9 is 0.1%. According to a strict interpretation of the legislation, waste from used LesEndo agar should not be categorized and managed as hazardous waste. However, evaporation has to be taken into consideration and the concentration may be above 0.1%. Therefore my evaluation is that waste from used LesEndo agar should be categorized and managed as hazardous waste to protect humans and the environment from being harmed.

Memoria para optar al Título Profesional de Profesional de Ingeniero Agrónomo / Comúnmente las plantas de alstroemeria (Alstromeria L.) son propagadas vegetativamente por división de su rizoma, pero este proceso consume tiempo y contribuye a la propagación de enfermedades. Es por esto que la mayoría de los híbridos de alstroemeria hoy en día son micro propagados in vitro mediante división de sus rizomas. La micropropagación es un método eficiente, limpio y prolijo. El objetivo de este estudio fue evaluar diferentes medios de cultivo con distintas concentraciones de agar (0,0; 3,5; 7,0 g•L-1) y citoquininas en forma de BAP (6-Benzylaminopurine) (0,0; 0,5; 1,0 y 2,0 mg•L-1) con el fin de determinar las mejores condiciones de estos factores para desarrollar un método eficiente de micropropagación para Alstroemeria pallida Graham. Se evaluó el peso del explantes (g), el número de brotes del rizoma y el largo de estos (cm), la longitud del rizoma (cm), el número de rizomas obtenidos y las tasas de proliferación, de mortalidad y de contaminación, también se hizo un registro fotográfico de los explantes.

A formação de biofilmes em superfícies que entram em contato com alimentos pode resultar em contaminação em qualquer parte do processo produtivo, podendo assim, ser causa de doenças transmitidas por alimentos. A formação de biofilmes por Staphylococcus ssp. é uma das grandes preocupações na indústria de lácteos, e também na área da medicina veterinária. A presente pesquisa teve como objetivo avaliar a capacidade de formação de biofilmes de Staphylococcus spp. isolados de laticínios produtores de queijo Minas Frescal e de leite de vacas com mastite subclínica. O estudo foi realizado em 3 fases, sendo a primeira uma caracterização fenotípica de 150 isolados de Staphylococcus spp. pelo teste do Ágar Vermelho Congo (AVC). Na segunda fase, foi realizada uma caracterização genotípica dos isolados de Staphylococcus spp., pesquisando os genes icaA, icaD, bap, bbp, cna, ebpS, eno, fib, fnbA, fnbB, clfA e clfB. Na terceira fase foram selecionadas 10 cepas com diferentes perfis genotípicos e três cepas padrão de Staphylococcus spp., para avaliação da capacidade de formação de biofilmes. Foi avaliada a capacidade de formação de biofilmes das 13 cepas em três tempos (12h, 48h e 96h), duas temperaturas (5°C e 25°C) e duas superfícies de contato (aço inoxidável 304 e polipropileno) tendo como primeira variável resposta a densidade óptica a 600 nm obtida utilizando metodologia do cristal violeta e a segunda variável resposta foi a contagem microbiológica de células viáveis (log10UFC cm-2) aderidas nas superfícies testadas. Foi utilizado um Delineamento Inteiramente Casualizado (DIC) com fatorial de 13x3x2x2. No AVC 38,7% (n=58) dos 150 isolados foram positivos para formação de biofilme. Observou-se que 93,3% (n=140) dos 150 isolados possuíam o gene clfA, 87,3% (n=131) o eno, 62,7% (n=94) o ebpS, 54,7% (n=82) o fib, 54% (n=81) o fnbA, 53,3% (n=80) o icaD, 47,3% (n=71) os genes icaA e clfB, 43,3% (n=65) o fnbB, 17,3% (n=26) o bap, 8% (n=8) o cna e 4% (n=40) o gene bbp. Pelo Teste de Friedman (&alpha;=0,05) os fatores cepa, temperatura e superfície foram significativos (p<0,0001) para as duas variáveis resposta na formação de biofilmes, enquanto que, entre os tempos de avaliação não houve diferença significativa (p>0,05). A maior produção de biofilme (avaliada por densidade óptica) foi observada a 25°C no aço inoxidável, sendo que as cepas S. epidermidis ATCC 35984 e S. aureus 119 foram as cepas que apresentaram maior formação de biofilme. Nas superfícies testadas foram observadas contagens microbiológicas na faixa entre 6,5 e 7,6 log10UFC cm-2 que sugerem formação de biofilme. Foram selecionadas 6 cepas para realizar a Microscopia Eletrônica de Varredura (MEV), as quais confirmaram a formação de biofilme em aço inoxidável e polipropileno em 5°C e 25°C sendo avaliadas á 12 e 96 horas. / The biofilm formation on surfaces, which are contact with food, can result in contamination in any part of the food processing, causing foodborne illness. The biofilm formation by Staphylococcus ssp. is one of the main concern in the dairy industry, and in the veterinary medicine field. This research aimed to evaluate the biofilm formation capacity of Staphylococcus spp., isolated from dairy producers of \"Minas Frescal\" cheese and cows with subclinical mastitis. The study was conducted in three stages, being the first a phenotypic characterization of 150 Staphylococcus spp. strains using the Congo Red Agar (AVC) test. In the second stage, the genotypic characterization with the screening of the gens icaA, icaD, bap, bbp, cna, ebpS, eno, fib, fnbA, fnbB, cIfA and clfB was performed. In the third stage, 10 strains with different genotypic profiles and 3 standard strains of Staphylococcus spp. were selected to evaluate the biofilm formation capacity. The biofilm formation capacity of 13 strains at three times (12h, 48h and 96h), two temperatures (5 °C and 25 °C) and two contact surfaces (stainless steel 304 and polypropylene) was evaluated. As a result, the optical density at 600 nm obtained by the methodology of crystal violet and the microbiological count of viable cells (log10UFC cm-2) (adhered to the tested surfaces) was analyzed. A completely randomized design was used in a factorial 13x3x2x2. In AVC agar, 38.7% (n = 58) of the 150 isolates were positive for biofilm formation. The 93.3% (n = 140) of the 150 isolates had the cIfA gene, 87.3% (n = 131) the eno, 62.7% (n = 94) the ebpS, 54.7% ( n = 82) fib, 54% (n = 81) the fnbA, 53.3% (n = 80) the icaD, 47.3% (n = 71) icaA and clfB gens, 43.3% (n = 65) the fnbB, 17.3% (n = 26) bap, 8% (n = 8) the cna and 4% (n = 40) bbp gene. The factors strain, temperature and surface were significant (Friedman test p <0.0001) for both response variables in the formation of biofilms, while, time did not have significant difference (p> 0.05). The increased production of biofilm (assessed by optical density) was observed at 25 °C in stainless steel surfaces. The strains S. epidermidis ATCC 35984 and S. aureus 119 were the strains that showed the highest production of biofilm. In the tested surfaces, microbiological counts were observed in the range between 6.5 and 7.6 log10CFU m-2 suggesting biofilm formation. The formation of biofilms in 6 evaluated strains has been confirmed by Scanning Electron Microscopy (SEM) on stainless steel and polypropylene on 5°C and 25°C with 12 and 96 h of incubation.

Diagnosis techniques and treatments for Escherichia coli mastitis in the dairy bovine were evaluated in two experiments. The first experiment evaluated eosin methylene blue agar as a method of distinguishing E.coli from other gram-negative mastitis pathogens. Escherichia coli will usually produce a green metallic sheen on eosin methylene blue agar. One hundred and twenty-nine milk samples or gram-negative isolates from milk samples were used to compare eosin methylene blue agar to a commercial biochemical test strip (the accepted standard). There was an intermethod agreement of 96.9% and a k-value of 93.7% indicating excellent agreement beyond chance between test methods. Eosin methylene blue agar is a reliable method for differentiation of E. coli from other gram-negative mastitis pathogens. The second experiment evaluated the efficacy of frequent milk-out as a treatment for E. coli mastitis. Sixteen Holstein dairy cows were divided into 2 blocks and randomly assigned to 1 of 4 treatment groups: 1) non-infected, not frequently milked-out, i.e. not treated (NI-NT), 2) experimentally infected with E. coli, not treated (EC-NT), 3) non-infected, frequently milked-out (NI-FMO), and 4) experimentally infected with E. coli, frequently milked-out (EC-FMO). Hours to bacterial, clinical and systemic cure were not different between the EC-NT and EC-FMO treatment groups. Serum a-lactalbumin concentrations were evaluated between treatment groups as a measure of udder health. Serum a-lactalbumin concentrations were higher in cows in the EC-NT treatment group than cows in the NI-NT, NI-FMO and EC-FMO treatment groups at 12 hours post-experimental challenge. Serum a-lactalbumin concentrations were higher in cows in the NI-FMO treatment group than in cows in the NI-NT, EC-NT and EC-FMO treatment groups at 36 hours post-experimental challenge. Results from this study do not support frequent milk-out as a treatment for E. coli mastitis. / Master of Science

Symbiotic dinoflagellates in the genus Symbiodinium are critical to the success of scleractinian reef corals in shallow tropical seas. These symbionts are commonly isolated from hosts and cultured separately in liquid media (f/2 or ASP8a), but initial isolations can be prone to abundant contaminants that can persist long-term in culture. To help remove these contaminants, I developed a solid growth substrate composed of 1% f/2 medium in agar, supplemented with a variety of antibiotics, to help isolate individual clones and establish new ?axenic? cultures. I found that an antibiotic cocktail of kanamycin (50 µg/mL), ampicillin (100 µg/mL) and streptomycin (50 µg/mL) was the most effective at eliminating visual signs of contamination without apparent harm to a variety of Symbiodinium in culture. Photophysiological measurements of Symbiodinium grown on f/2 agar plates, taken using an Imaging Pulse Amplitude Modulated (I-PAM) fluorometer, were comparable with those grown in liquid f/2, both with and without antibiotics. Eight types of Symbiodinium in clades A-D grown on f/2 agar plates at low irradiance (19-46 µmol photons m-2 s-1) were exposed to higher irradiance conditions (50-90 µmol photons m-2s-1) for 13 days and their photosynthetic efficiencies (Fv/Fm) were compared using the I-PAM. There were significant differences among and within clades, except for two types in clade C (C2 and C3) which did not perform differently from eachother. All types showed reduced Fv/Fm after 12 days in higher light. Type D1a showed high mortality after 13 days of higher light stress. Finally, preliminary work to fluorescently label Symbiodinium determined that available vital stains were not generally practical for symbiosis studies. Attempts to transform Symbiodinium with a variety of plasmids containing fluorescent reporters and/or genes for antibiotic resistance were not successful, but did provide a baseline for future work. In summary, Symbiodinium cultures grown on solid substrates supplemented with antibiotics are useful for: (1) isolating individual cells or clones for subsequent applications and establishing and maintaining ?axenic? cultures that are free of observable contaminants; (2) directly comparing the photophysiology of different cultures using an I-PAM fluorometer; (3) quantifying cells on agar plates using the I-PAM and (4) selecting possible transgenic symbionts for symbiosis studies.

Quorum sensing (QS) is a type of microbial communication used by bacteria to coordinate their behavior based on population density, regulating complex processes like biofilm formation and virulence, among other behaviors. Quorum quenching (QQ), on the other hand, disrupts this communication, usually by degradation of the QS signaling molecule. QQ offers a potential strategy for controlling bacterial behaviors linked to pathogenicity and biofouling. Despite significant advances in understanding and modeling the spatial-temporal behavior of QS, predictive modeling of QQ remains nascent, with a notable gap in the quantitative assessment of QQ's impact on QS. Here we show quantitative evaluation and characterization of the effect of QQ on QS in agar-based experiments, combined with an experimentally validated computational model. This research utilizes green fluorescence in E. coli MG 1655 as an indicator of QS activation, focusing on the degradation of Acyl-Homoserine Lactone (AHL), a key QS molecule in Gram-negative bacteria linked to pathogenicity, by the AiiA enzyme in engineered AiiA-producing Salmonella Typhimurium 14028. Our findings suggest that QQ more effectively influences QS in spatial configurations of the populations with larger interaction surfaces and shorter diffusion distances. Contrary to our initially held hypothesis, the primary effect of QQ is not a delay in QS onset but rather an attenuation of QS activity, with the area-under-the-curve of fluorescence serving as a quantitative metric. This study also introduces, to the best of our knowledge, one of the first instances of experimentally validated predictive modeling for QQ, applied to agar-based experimental setups. We posit that the quantitative experimental characterization and modeling framework presented in this research will enhance the understanding of bacterial community interactions. Enhanced comprehension of QQ and QS behaviors holds significant promise for advancing practical applications, particularly in mitigating or diminishing undesirable QS-associated activities. This is especially relevant in areas like biofouling, waste treatment, and the reduction of infections and progression of diseases in plants and animals, areas increasingly important as concerns about drug resistance in microbes and food security escalates. / Master of Science / One of the ways bacteria communicate with each other is called quorum sensing (QS), where they use chemical signals to organize and time group behavior, including forming communities encapsulated in protective layers, called biofilms, and engaging in virulent attacks against hosts. Quorum quenching (QQ) in bacteria, however, disrupts this communication system, usually by breaking down the chemical signals that bacteria use to send messages to each other. Even though QS has been studied extensively, determining how to predict and control QQ is still a nascent area of research. Here, we studied and characterized how QQ affects QS by doing experiments with bacteria populations in agar (a jelly-like substance) and applied a computational model to explain and ultimately predict the experimental observations. Engineered QS population (E. coli MG 1655) produced Acyl-Homoserine Lactone (AHL) signaling molecules, and engineered QQ bacteria (S. Tm 14028) used the Autoinducer Inactivation A (AiiA) enzyme to break down the AHL. According to our results, QQ doesn't delay the QS bacteria's group behaviors (in our case, green fluorescent signal production); it weakens the signal instead. Understanding QQ and QS better, especially through measurements and modeling, could lead to expanded methods of deterring harmful bacterial behavior, managing waste better, and stopping diseases in plants, animals, and humans, especially with the concerning rise of drug-resistant microbes and food security. One exciting possibility is using QQ to protect plants from bacterial infections. This could be a way to shield our crops without always relying on antibiotics.

Vätskekromatografi är vanligt förekommande metod för separationer av molekyler inom bioteknik. Det finns inom begreppet vätskekromatografi många olika tekniker och vilken specifik teknik som är mest lämplig för en given separation beror på egenskaperna hos den avsedda målmolekylen. Egenskaper för vilka målmolekyler kan separeras är bland annat storlek, laddning och affinitetsintreaktioner. Porositeten är i den stationära fasen en avgörande egenskap, eftersom porernas tillgänglighet påverkar den stationära fasens tillgängliga yta. Således är förmågan att bestämma porositeten en avgörande aspekt vid tillverkningen av den stationära fasen. Agaros utgör ofta den stationära fasen, på grund av modifieringsmöjligheterna hos polysackaridkedjorna.  Bio-Works Workbeads 40/1000 SEC är agarbaserade pärlor för den stationär fas i storleksutslutningskromatografi. Pärlorna är tvärbundna två gånger för att få tillräcklig styvhet och för att uppnå en önskad fördelningskoefficient. Det första tvärbindningssteget bestämmer strukturen på pärlorna och därmed porositeten, medan det andra tvärbindningssteget försäkrar att pärlorna får tillräcklig styvhet. För produkten Workbeads 40/1000 är de önskade fördelningskoefficienterna mellan 0,2-0,35 för thyroglobulin, 0,62-0,75 för albumin (från äggvita) och 0,85-0,95 för ribonukleas A. För att bestämma förhållandet mellan koncentrationerna av reaktanterna NaOH och allylbromid i den första tvärbindningen och de resulterande egenskaperna hos den stationära fasen (fördelningskoefficienter, styvhet och porstorlek) ändrades koncentrationerna av reaktanterna i den första tvärbindningen. Samtliga resulterande pärlorna med de ändrade koncentrationerna analyserades med hjälp av en ÄKTAexplorer. Pärlornas styvhet bestämdes genom att analysera kraschflödet i den packade bädden. Det fanns inga tecken på några skillnader mellan de tillverkade pärlproverna. De icke-specifika interaktionerna analyserades genom att jämföra fördelningskoefficienterna i en standard PBS-buffert med en hög saltkoncentrationsbuffert. Polaritetsförändringen i den mobila fasen kan påverka de icke-specifika interaktionerna eftersom hydrofobiska interaktioner då främjas. Fördelningskoefficienternas kurvor jämfördes också för att analysera likheter och skillnader för olika proteiner. Skillnader i formen på fördelningskoefficientkurvorna kan innebära att porstorleken inte är den enda bidragsfaktorn till skillnaderna i fördelningskoefficienterna. Därför kan skillnaden innebära effekter av icke-specifika interaktioner och det fanns det indikationer på att vissa icke-specifika interaktioner uppstod mellan proteinerna och agarpärlorna. Det finns dock ingen skillnad mellan de pärlor med många tvärbindningar och de med få tvärbindningar. Porstorleken och porstorleksdistribution bestämdes genom att använda en uppsättning dextran med inverterad storleksutslutningskromatografi.  Efter varje tvärbindningsreaktioner bestämdes antalet bildade tvärbindningar genom titrering med silvernitrat. Det visade sig att koncentrationen av allylbromid hade en större inverkan på de resulterande egenskaperna jämfört med koncentrationen av NaOH. En ökad koncentration av allylbromid visade på ett ökat antal bildade tvärbindningar. Experiment med många bildade tvärbindningar visade sig ha en större porstorlek och högre fördelningskoefficienten jämfört med experiment med lägre mängd bildade tvärbindningar. / Liquid chromatography is a method that is extensively used for separations in biotechnology at present. There are many different techniques within the concept of liquid chromatography and which specific technique that is most suitable depends on the properties of the intended target molecule. Properties for which target molecules can be separated are among others, size, charge, and affinity interactions. The porosity is a crucial property of the chromatographic resin, as the accessibility of pores determines the accessible surface area of the resin. Thus, the ability to determine the porosity is a crucial aspect in the manufacturing of the resin. Agarose often constitutes the chromatographic resin, due to the modification possibilities of the polysaccharide chains.  Bio-Works Workbeads 40/1000 SEC are agar-based size exclusion chromatography beads. The beads are cross-linked twice in order to obtain sufficient rigidity and to achieve a desired distribution coefficient. The first cross-linking step determines the structure of the beads thereby stets the porosity and the second cross-linking step ensures a sufficinet rigidity. For the product Workbeads 40/1000 the desired distribution coefficients is between 0.2-0.35 for thyroglobulin, 0.62-0.75 for albumin (from egg white) and 0.85-0.95 for ribonuclease A. To determine the relation between the concentrations of two reactants NaOH and allyl bromide in the first cross-linking step and the resulting properties of the resin (distribution coefficients, rigidity, and pore size), the concentrations in the first cross-linking were changed. All resulting beads with the altered reactant concentrations were analyzed using an ÄKTAexplorer. The rigidity of the beads was determined by analyzing the crash flow rate of the packed bed. There were no indications of any differences between the manufactured resin samples. The non-specific interaction was analyzed by comparing the distribution coefficients in a standard PBS buffer to a high salt concentration buffer. The polarity change in the mobile phase might affect the non-specific interactions since hydrophobic interactions would be promoted. The distribution coefficient curves were also compared for similarities and differences between different proteins. Differences in the shape of the distribution coefficient curves might imply that the pore size is not the only contribution factor to the distribution coefficient differences. Hence, the difference might imply the effect of non-specific interactions. It was indicated that some non-specific interactions occurred between the proteins and the agar beads. However, there was no difference between beads of varying cross-linking degree. The porosity and pore size distribution were determined by utilizing a set of dextran’s with Inverse Size Exclusion Chromatography (ISEC). After each cross-linking step the number of cross-links was determined by titration with silver nitrate (AgNO3). It was shown that the concentration of allyl bromide had a greater impact on the resulting properties compared to the concentration of NaOH. An increased concentration of allyl bromide showed increased number of formed cross-links. Experiments with many formed cross-links was shown to have a larger pore size and higher distribution coefficient Kd compared to experiments with lower amount of formed cross-links.

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

O aquecimento climático apresenta um impacto sem precedentes na biodiversidade, afetando o desempenho fisiológico e a tolerância dos organismos, particularmente para ectotermos terrestres, que funcionam perto dos seus limites fisiológicos superiores. No entanto, estas relações entre clima e biodiversidade têm sido estudadas principalmente em grandes escalas que não explicam o microclima, entendido como clima numa escala individual. Atualmente o microclima é considerado essencial para compreender o impacto de suas alterações sobre a diversidade biológica com interesses focais sobre dado táxon. A Mata Atlântica do Brasil apresenta uma diversidade fisionômica e biológica que são causa e efeito do clima. Essas relações amplas não são suficientes para explicar as opções de termo e hidro regulação disponíveis para pequenos animais, que é particularmente importante para anfíbios anuros, um táxon particularmente diversificado. Portanto, o objetivo deste projeto foi investigar os microclimas que os anuros da Mata Atlântica podem experenciar em diferentes fisionomias, estratos e habitats fragmentados (áreas abertas, borda e interior da floresta). Todos esses fatores devem influenciar variação hidrotermal numa escala espaço temporal, compatível com o tamanho do corpo dos adultos. Para definir estas relações, usamos um conjunto de indicadores (modelos de ágar com sensores e imagens infravermelhas) que estimam a temperatura operativa e disponibilidade de água no ambiente. Este estudo foi baseado em dados de tamanho corporal real que incluem a maioria dos gamas de tamanhos e permeabilidade de pele de anuros presentes no Parque Estadual Intervales. Usamos como unidade amostral pares de modelos de ágar que simulam a forma e extremos de tamanho corpóreo (pequena vs. grande) e tipos de permeabilidade (totalmente permeáveis vs. impermeáveis). Em geral, modelos permeáveis, especialmente de tamanhos menores, apresentam maior perda de água em áreas de vegetação mais baixa, especialmente no dossel, áreas abertas e em mata de altitude. Os dados obtidos permitiram propor hipóteses de impacto diferencial de altas e baixas temperaturas e baixa umidade relativa, de acordo com a faixa microambiental ocupada pelos anuros. Estas informações ajudam no entendimento da paisagem hidrotermal e do impacto das mudanças climáticas sobre a história natural, em interpretação de dados eco-fisiológicos, modelagem climática e previsão do impacto do clima sobre anuros, particularmente no contexto da dinâmica térmica e do balanço hídrico em escalas espaciais e temporais / Climate warming presents an unprecedented impact to global biodiversity, affecting physiological performance and tolerance of organisms, particularly for terrestrial ectotherms, which function near their upper physiological limits. However, the relationships between climate and biodiversity have been studied mainly at large scales that do not explain microclimate, understood as climate at individual scale. Microclimate is currently considered essential to understand the impact of climate on biological diversity with focal interests on given taxa. Atlantic Forest of Brazil presents physiognomic and biological diversity that are cause and effect of climate. These relationships are not enough to explain options of thermo and hydro-regulation available for small animals, particularly important for anuran amphibians, a particularly diverse taxon. Therefore, the aim of this project was to investigate the microclimates from Atlantic Forest experimented by anurans in different physiognomies, stratums and fragmented habitats (open areas, edge and inside forest). All these factors should influence hydrothermal variation at temporal-space scale compatible with the body size of adults. To define this diversity, we used a set of indicators (agar models with sensors and infrared images) that estimate the operative temperature and water availability. This study was based on actual body size data to include most ranges of size and of skin\'s permeability. We used as sampling unit; pairs of agar\'s models that simulate shape and two body size extremes (small vs. large) and two types of permeability (fully permeable vs. impermeable). In general, permeable models especially of smaller sizes have a higher water loss in lower vegetation areas, especially in Canopy, Open Area and High-Altitude Forest. The data collected allowed to propose hypotheses regarding differential impact of high and low temperatures, and low relative humidity, according to the micro-environmental range occupied by anurans. This information will help to understand hydrothermal landscapes and impact of climate on natural history, interpretation of eco-physiological data, climate modeling and prediction of the impact of climate change on anurans, particularly in the context of thermal dynamics and water balance in spatial and temporal scales